GSM6025649: tonsillar.CD4.CD57-.VDJ; Homo sapiens; OTHER
Children with routine tonsillectomy were recruited for tonsils collection, and their blood may be also obtained. After mechanical disruption by surgical blades, tonsillar lymphocytes were collected by gradient centrifugation using Ficoll-Paque Plus tubes (GE Healthcare), and the PBMCs were also collected by gradient centrifugation. CD57+ CD4+ T cells (7AAD- CD19- CD3+ CD4+ CD8- CD57+) and CD57- CD4+ T cells (7AAD- CD19- CD3+ CD4+ CD8- CD57-) from tonsil and blood samples of a child were FACS-purified into 1.5-ml Eppendorf tubes with FACS buffer. Single cell gene expression and VDJ libraries were prepared by Biomolecular Resource Facility at ANU using Chromium Next GEM Single Cell 5' Kit v2 Dual Index and Single Cell Human TCR Amplification Kit (10x Genomics) according to the manufacturer's instructions. The library was sequenced in S1 flow cell with 2 x 50 bp paired-end reads by NovaSeq 6000 sequencer (Illumina).